PROJECT SUMMARY As with most synthetic chemists, CoALA Biosciences is inspired by naturally-occurring polyketide assembly lines, which synthesize diverse medicines through reactions that generate carbon-carbon bonds and precisely set stereocenters. If chemists were to collaborate with the enzymes in these assembly lines, syntheses of compounds in the drug discovery process would be greatly accelerated and yield larger quantities for preclinical and clinical studies. CoALA Biosciences will distribute these enzymes and those from related pathways so that chemists can employ these biosynthetic biocatalysts. They will be sold in easy-to-use kits so that scientists who have never handled an enzyme only need to add a mixture from a single container to a solution of their substrate. Since most chemists would prefer to directly purchase the organic fragments needed in the synthesis of a larger molecule, chiral building blocks up to seven carbons long and containing up to four contiguous stereocenters will be produced through chemoenzymatic synthesis at CoALA Biosciences and made available on the gram scale (Specific Aim 1). These chiral building blocks, not available from other commercial sources, will be a welcome addition to the limited set of chiral precursors abundant enough to be utilized in a synthesis. Other biocatalysis companies (e.g., ChiralVision, Codexis, Discovery Biocatalysts, Enzymicals, Iris Biotech, Prozomix, SyncoZymes) empower specific chemical transformations but do not offer enzymes from assembly lines evolved to set multiple stereocenters on carbon chains or facilitate the multistep synthesis of organic compounds. By refining the techniques employed to generate diketide and triketide chiral building blocks, CoALA Biosciences will seek to access tetraketide and pentaketide fragments during Phase II. A central mission of the company is to make polyketides customizable like oligonucleotides and polypeptides. CoALA Biosciences will also empower hundreds of laboratories studying natural product biosynthesis by offering a variety of relevant enzymes and enzyme kits (Specific Aim 2). Thus, similar to how enzymatic tools from New England Biolabs can be utilized to engineer DNA, versatile enzymes from CoALA Biosciences can be employed to precisely manipulate polyketides and study the biosynthesis of natural products.